LOCUS       sequence-417740-        9202 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian overexpression vector for C-terminally TwinStrep and His 
            tagged human Ttyh1 (W417R).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9202)
  AUTHORS   Bell TA, Luce BE, Hakim P, Ananda VY, Dardari H, Nguyen TH, 
            Monshizadeh A, Chao LH
  TITLE     Prominin 1 and Tweety Homology 1 both induce extracellular vesicle 
            formation.
  JOURNAL   Elife. 2024 Aug 13;13:e100061. doi: 10.7554/eLife.100061.
  PUBMED    39168849
REFERENCE   2  (bases 1 to 9202)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2024-08-13"; volume: "13:e100061. doi"; pages: " 
            10.7554/eLife.100061"
COMMENT     Sequence Label: pLX304-HsTtyh1-TwinStrep-His
FEATURES             Location/Qualifiers
     source          1..9202
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        66..369
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        370..573
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     524..544
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    596..620
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2001..2021
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(2034..2039,2040..2063,2064..2099,2100..2123)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     CDS             2139..2168
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     misc_feature    2210..2798
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2263..2283)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2681..2692)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2870..3103
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3181..3302
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3218..3237)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3272..3291
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3342..3477
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3497..3516)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3498..3516)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3526..3543)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3526..3542)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3535..3557)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3684..4139
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3771..3790)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3981..4002
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4165..4269
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4270..4338,4339..5130)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4488..4507)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5301..5889
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5790..5809
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6043..6060
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6177..6198
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6213..6218,6219..6236,6237..6243)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6251..6267
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6256..6278
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6275..6291
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6275..6291
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6310..6330
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6312..6330
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6358..6584
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6585..6765
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6812..6937
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7430..7663
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7848..7892
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        8067..8567
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     8486..8504
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             8579..8974
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9034..9151
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gcccggggtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
       61 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      121 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      181 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      241 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      301 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      361 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      421 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      481 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      541 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactgtcgg gatcaacaag
      601 tttgtacaaa aaagcaggct atgggggcgc ccccgggcta ccggccctca gcttgggtgc
      661 atctcctcca ccagctgccc cgcgccgact tccagctccg cccggtgccc agcgttttcg
      721 cgccccaaga gcaggaatac cagcaggcct tgttgctggt ggcggccttg gcgggcctgg
      781 gcttgggcct gagcctcatt ttcatcgctg tctacctcat ccgcttctgc tgctgccggc
      841 cccccgagcc ccccgggtcc aagatcccct cgcccggggg aggctgcgtc acctggagct
      901 gcattgtcgc ccttctcgcc ggctgcactg gcattggcat cggtttctat ggcaacagtg
      961 agaccagtga tggggtgtcc cagctcagct ctgcgctgct gcacgccaac cacacactca
     1021 gcaccattga ccacctggtg ttggagacgg tggagaggct gggcgaggcg gtgaggacag
     1081 agctgaccac cctggaggag gtgctcgagc cgcgcacgga gctggtggct gccgcccgag
     1141 gggctcgacg gcaggcggag gctgcggccc agcagctgca ggggctggcc ttctggcagg
     1201 gagtgcccct gagccccctg caggtggctg aaaatgtgtc ctttgtggag gagtacaggt
     1261 ggctggccta cgtcctcctg ctgctcctgg agctgctggt ctgcctcttc accctcctgg
     1321 gcctggcgaa gcagagcaag tggctggtga tcgtgatgac agtcatgagt ctcctggttc
     1381 tcgtcctgag ctggggctcc atgggcctgg aggcagccac ggccgtgggc ctcagtgact
     1441 tctgctccaa tccagaccct tatgttctga acctgaccca ggaggagaca gggctcagct
     1501 cagacatcct gagctattat ctcctctgca accgggccgt ctccaacccc ttccaacaga
     1561 ggctgactct gtcccagcga gctctggcca acatccactc ccagctgctg ggcctggagc
     1621 gagaagctgt gcctcagttc ccttcagcgc agaagcctct gctgtccttg gaggagactc
     1681 tgaatgtgac agaaggaaat ttccaccagt tggtggcact gctacactgc cgcagcctgc
     1741 acaaggacta tggtgcagcc ctgcggggcc tgtgcgaaga cgccctggaa ggcctgctct
     1801 tcctgctact cttctccctg ctgtctgcag gagcgctggc cactgccctc tgcagcctgc
     1861 cccgagcccg ggccctcttc ccacccagga atccaagcgc tttgtgcagt ggcagtcgtc
     1921 tatctgagcc cctcctcccg gctggactgg agcctggctc ccctcttcgt tccttccctg
     1981 gctgccggag gagaccccac gagaatttgt actttcaagg cggtggagga gggagtgcct
     2041 ggtctcatcc acagtttgaa aagggaggag gatccggagg tggtagtggc ggatctgcat
     2101 ggtcccaccc tcagtttgag aagggcggcg gaggttctca tcaccaccac catcatcatc
     2161 accaccacta ataatgagct agcgctaacc ggtggcgcgt taagtcgaca atcaacctct
     2221 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2281 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2341 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2401 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2461 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     2521 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2581 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     2641 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     2701 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     2761 tcagacgagt cggatctccc tttgggccgc ctccccgcgt cgactttaag accaatgact
     2821 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     2881 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
     2941 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     3001 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     3061 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtacgta tagtagttca
     3121 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     3181 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3241 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3301 tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc
     3361 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3421 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3481 ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
     3541 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     3601 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     3661 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3721 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3781 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3841 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3901 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3961 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4021 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4081 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     4141 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4201 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4261 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4321 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4381 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4441 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4501 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4561 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4621 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4681 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4741 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4801 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4861 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4921 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4981 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5041 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5101 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5161 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5221 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5281 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5341 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5401 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5461 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5521 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5581 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5641 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5701 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5761 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5821 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5881 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5941 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6001 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6061 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     6121 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     6181 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     6241 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     6301 gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt
     6361 agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct
     6421 tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg
     6481 ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc
     6541 attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag
     6601 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     6661 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     6721 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag
     6781 ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga
     6841 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
     6901 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
     6961 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
     7021 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
     7081 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
     7141 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
     7201 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
     7261 caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt
     7321 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
     7381 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
     7441 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg
     7501 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
     7561 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
     7621 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
     7681 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
     7741 tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca
     7801 caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag
     7861 aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc
     7921 tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt
     7981 ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga
     8041 cccacctccc aaccccgagg ggacccttgc gccttttcca aggcagccct gggtttgcgc
     8101 agggacgcgg ctgctctggg cgtggttccg ggaaacgcag cggcgccgac cctgggtctc
     8161 gcacattctt cacgtccgtt cgcagcgtca cccggatctt cgccgctacc cttgtgggcc
     8221 ccccggcgac gcttcctgct ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt
     8281 gccggacgtg acaaacggaa gccgcacgtc tcactagtac cctcgcagac ggacagcgcc
     8341 agggagcaat ggcagcgcgc cgaccgcgat gggctgtggc caatagcggc tgctcagcag
     8401 ggcgcgccga gagcagcggc cgggaagggg cggtgcggga ggcggggtgt ggggcggtag
     8461 tgtgggccct gttcctgccc gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt
     8521 cggcagtcgg ctccctcgtt gaccgaatca ccgacctctc tccccagggg gtaccaccat
     8581 ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa
     8641 cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg
     8701 catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt
     8761 gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa
     8821 tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca
     8881 tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg
     8941 tgaattgctg ccctctggtt atgtgtggga gggcctgcag ctgcagtagt aagaattcta
     9001 gatcttgaga caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg
     9061 gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt
     9121 acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc
     9181 actttggcgc cggctcgagg gg
//
